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Cow sex-control sperm freezing preservation seminal fluid dilution

A technology of cryopreservation and diluent, which is applied in the field of dairy cow reproduction, can solve the problems of short effective fertilization time, low vitality and acrosome integrity rate, and no patents have been seen, so as to improve the acrosome integrity rate of sperm and improve sperm quality. The effect of improving the activity rate and reducing the production cost

Inactive Publication Date: 2007-03-14
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method of separating semen also has some defects: the pregnancy rate of sex-separated sperm is lower than that of non-separated sperm; the time of effective fertilization ability of sex-separated sperm in the female reproductive tract is short; the motility and acrosome integrity rate after thawing are low , cows and other animals need to be inseminated close to the time of ovulation and in the deep part of the uterus (close to the fertilization site) to have a good effect
[0003] After searching, no patent similar to the present invention has been seen so far

Method used

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  • Cow sex-control sperm freezing preservation seminal fluid dilution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Preparation of liquid A (basic buffer): mix 20g / L tris (Tris), 15g / L citric acid, and 9g / L glucose, dilute to 1L with distilled water, and obtain the described liquid A ;

[0031] (2) Preparation of liquid B (basic dilution): first uniformly mix 20% of fresh egg yolk and 80% of liquid A, then add 1000IU / mL penicillin and 1000IU / mL streptomycin, and mix uniformly to obtain the described B liquid;

[0032] (3) Prepare liquid C (diluent 1): uniformly mix 95% liquid B and 5% pig semen to obtain liquid C;

[0033] (4) Preparation of liquid D (diluent 2): first uniformly mix 81% of liquid B and 5% pig semen, and then evenly mix with 14% of glycerin to obtain the liquid D;

[0034] (5) Dilute the sex-controlled semen of dairy cows with liquid C in step (3) to half of the total volume before centrifugation, and equilibrate at 4°C for 1.5 hours to obtain mixed solution 1;

[0035] (6) Add liquid D to the mixed liquid 1 in step (5) to 3 / 4 of the total volume before centrif...

Embodiment 2

[0047] (1) Preparation of liquid A (basic buffer): mix 26g / L tris (Tris), 10g / L citric acid, and 13g / L glucose, dilute to 1L with distilled water to obtain the described liquid A ;

[0048] (2) Preparation of liquid B (basic dilution): first uniformly mix 20% of fresh egg yolk and 80% of liquid A, then add 500IU / mL penicillin and 500IU / mL streptomycin, and mix uniformly to obtain the described B liquid;

[0049] (3) Preparation of liquid C (diluent 1): uniformly mix 90% of liquid B and 10% of pig semen to obtain liquid C;

[0050] (4) Preparation of liquid D (diluent 2): first uniformly mix 76% of liquid B and 10% of pig semen, and then evenly mix with 14% of glycerin to obtain the liquid D;

[0051] (5) Dilute the sex-controlled semen of dairy cows with liquid C in step (3) to half of the total volume before centrifugation, and equilibrate at 4°C for 1.5 hours to obtain mixed solution 1;

[0052] (6) Add liquid D to the mixed liquid 1 in step (5) to 3 / 4 of the total volume...

Embodiment 3

[0064] (1) Preparation of liquid A (basic buffer): mix 23g / L tris (Tris), 13g / L citric acid, and 11g / L glucose, dilute to 1L with distilled water to obtain the described liquid A ;

[0065] (2) Preparation of liquid B (basic dilution): first uniformly mix 20% of fresh egg yolk and 80% of liquid A, then add 500IU / mL penicillin and 500IU / mL streptomycin, and mix uniformly to obtain the described B liquid;

[0066] (3) Preparation of liquid C (diluent 1): uniformly mix 85% of liquid B and 15% of pig semen to obtain liquid C;

[0067] (4) Preparation of liquid D (diluent 2): firstly mix 71% of liquid B and 15% of pig semen evenly, and then evenly mix with 14% of glycerin to obtain the liquid D;

[0068] (5) Dilute the sex-controlled semen of dairy cows with liquid C in step (3) to half of the total volume before centrifugation, and equilibrate at 4°C for 1.5 hours to obtain mixed solution 1;

[0069] (6) Add liquid D to the mixed liquid 1 in step (5) to 3 / 4 of the total volume ...

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PUM

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Abstract

The invention relates to a method for freezing and storing diluted semen of cow. Wherein, the invention is characterized in that: preparing said diluted semen into A, B, C, D deals; mixing them with right ratio; adding 5-20% pig semen into C and D deals, to improve the semen acrosome integral rate of cow semen control liquid, and reduce the density of semen, to save cost. The experiment has proved that the semen survival rate of stored diluted semen can reach 50.88-62.88%, while the average number is 57.41%, which is 15.13% higher than traditional method, and the semen acrosome integral rate can reach 50.88-53.20%, while the average number is 51.90%, which is 7.0% higher than traditional method.

Description

technical field [0001] The invention belongs to the technical field of dairy cow reproduction, and in particular relates to the preparation of semen diluent for cryopreservation of sex-controlled sperm of dairy cows. technical background [0002] Since 1989, Johnson et al. reported for the first time that the live X sperm and Y sperm of rabbits were successfully separated by using the flow cytometry retrieval separator, and live offspring were produced after fertilization with the separated sperm, which made the sperm separation technology of the flow cytometry retrieval separator A breakthrough has been made in the research of (Johnson L A, Flook J P, Hawk H W.Sex prpmem selection in rpmabbits: live birpmth frpmmom X and Y spermpmmseparpmated by DNA and cell sorpmting. Biol RPMep rpmod, 1989, 41: 199-203). Since then, the use of this technology to separate active X sperm and Y sperm has been successfully applied to artificial insemination, in ...

Claims

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Application Information

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IPC IPC(8): A61K35/52A61K47/46A61K47/26A61K47/22A61K47/10
Inventor 杨利国桑雷
Owner HUAZHONG AGRI UNIV
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